The pathogenesis of B cell malignancies is a dynamic process
The pathogenesis of B cell malignancies is a dynamic process involving reciprocal interactions between malignant cells and surrounding components in the tumour microenvironment. In the case of certain B cell lymphomas, neoplastic cells migrate to lymphoid sites and prompt the infiltration of non-malignant immune cells. The infiltration of cytotoxic lymphocytes correlates with tumour cell killing and good prognosis. However, most immune cell types in the tumour niche often become unresponsive or end up promoting tumour growth.
Detailed workflow for successful sorting of six immune cell populations
This white paper and its associated webinar describe a detailed workflow for successful sorting of six immune cell populations from a tumour niche in a mouse model of B cell lymphoma. The study has uncovered strategies for increasing sample throughput. It has also demonstrated how to perform a detailed examination of critical cell populations in tumour responses using simultaneous analysis of proteins and mRNA transcripts at the single-cell level, especially for rare subsets.
The authors of this study examined a murine model of B cell lymphoma, focusing on the function of the B and T cell lymphocyte attenuator (BTLA) in the progression of the tumour. They designed a high-throughput sorting strategy that enabled the isolation of six cell populations from the tumour niche using the BD FACSymphony™ S6 Cell Sorter. Following this, they characterized the sorted cells using downstream multi-omics analysis on the BD Rhapsody™ system.
The loss of BTLA function may increase the proportion of CD8+ T cells
From the data obtained, the authors concluded that the loss of BTLA function may increase the proportion of CD8+ T cells expressing CD279 (PD-1), a protein that plays a key role in tumorigenesis by modulating T cell immune response. The authors observed this both in endogenous CD8+ cells, as well as those that were transferred to the mice. The analysis also showed that the endogenous cells expressed several proteins and mRNA transcripts like CD137 and Gzma, suggesting that they had undergone activation. These activated cells could potentially contribute to eliminating the tumour cells. The authors propose that further analysis of the sorted populations could shed light on the interplay of different cell types in B cell lymphoma.
The webinar associated with the study also discusses the following:
- Steps for cell preparation before 6-way sorting to reduce cell stress and cell loss
- Strategies to increase sample throughput by enabling sample multiplexing and concurrent multiparametric analyses of surface proteins and mRNA transcripts
- Exploiting Uniform Manifold Approximation and Projection (UMAP) and other easy-to-use utility tools for demultiplexing samples and deep single-cell analysis
Disclaimer: The work described in this white paper is an advertisement and has not been peer-reviewed.
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